Evidence Source
| Description | Cleavage Assaysâ€â€In vitro native substrate cleavage assays performed at enzyme to substrate ratios of 1:10 (w/w) of chemokines, vimentin, IGFBP-7, cystatin C, galectin-1, and secreted protein, acidic and rich in cysteine (SPARC) by sMT6- MMPdeltaF were performed in a 10-µl reaction containing 50 mM HEPES, 200 mM NaCl, 5 mM CaCl2, pH 7.4 for 16 h at 37 °C. Chemokine cleavage assay products were analyzed by MALDI-TOF mass spectrometry (MS) on a Voyager-DE STR (Applied Biosystems) using sinapinic acid matrix (33) and confirmed by silver-stained 15% Tris-Tricine SDS-PAGE. Chemokine cleavage was defined to be positive when the mass spectrometry spectra showed a cleavage product with ~20% ion intensity of the full-length chemokine. |
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Method |
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Experimental system |
cell free |
Perturbation |
none |
Protease inhibitor(s) used |
|
Directness |
direct |
Physiological relevance |
unknown |
Evidence Code(s) |
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Database / Laboratory |
- Lab: Overall Lab |
Raw data repository |
Evidence for:
0
N-termini
0
C-termini
Publication(s) |
Biochemical characterization and N-terminomics analysis of leukolysin, the membrane-type 6 matrix metalloprotease (MMP25): chemokine and vimentin cleavages enhance cell migration and macrophage phagocytic activities. Starr AE, Bellac CL, Dufour A, Goebeler V, Overall CM J Biol Chem. 2012 Apr 13;287(16):13382-95. doi: 10.1074/jbc.M111.314179. Epub 2012 Feb 24. PMID: 22367194 |
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